陈琨,吴海铃,杨创业,王庆恒

• 1:广东海洋大学水产学院
• 2:广西科学院广西海洋科学院(广西红树林研究中心)
• 3:广东省珍珠养殖与加工工程技术研究中心
• 4:广东省珍珠科技创新中心

摘要(Abstract)：

【目的】探究急性低氧胁迫对马氏珠母贝（Pinctada fucata martensii）生长、免疫及矿化相关基因表达的影响，为马氏珠母贝抗逆品系选育提供参考。【方法】将马氏珠母贝从常氧条件下移至2 mg·L-1低溶解氧水体，比较其在低氧胁迫0、12、24和48 h时免疫与抗氧化相关基因（caspase-3、HSP90、CAT和SOD）、生长相关基因（EGFR、FGF18、OSR1和tgfbr1）和矿化相关基因（pif177、aspein、nacrein和TIMP）的表达水平。【结果】急性低氧胁迫后，HSP90的表达水平12 h时低于对照组（P <0.05）,caspase-3和CAT的表达在胁迫24 h时显著增加（P <0.05），而SOD的表达无明显变化（P> 0.05）；矿化相关基因pif177、aspein、nacrein在急性低氧胁迫后的表达均显著降低，TIMP仅在12 h时显著降低（P <0.05）；生长相关基因EGFR、FGF18和OSR1基因的表达也出现显著下降（P <0.05）,tgfbr1的表达在24 h时显著高于对照组（P <0.05）。【结论】急性低氧胁迫上调免疫与抗氧化相关基因中caspase-3、CAT的表达，抑制HSP90的表达，抑制矿化相关基因的表达，抑制生长相关基因中EGFR、FGF18和OSR1的表达，上调tgfbr1的表达。

关键词(KeyWords)： 马氏珠母贝;基因表达;急性低氧胁迫

Abstract:

Keywords:

基金项目(Foundation): 国家贝类产业技术体系（CARS-49）;; 广东省普通高校重点领域专项（2020ZDZX1045）;; 广东省现代农业产业技术体系贝藻类产业创新团队（KJ146）

作者(Author): 陈琨,吴海铃,杨创业,王庆恒

参考文献(References)：

• [1] WEI H, HE Y C, LI Q J, et al. Summer hypoxia adjacent to the Changjiang Estuary[J]. Journal of Marine Systems,2007, 67(3/4):292-303.
• [2] VAQUER-SUNYER R, DUARTE C M. Thresholds of hypoxia for marine biodiversity[J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(40):15452-15457.
• [3]张文斌,吕振波,张莹,等.缺氧胁迫对菲律宾蛤仔(Ruditapes philippinarum)生理代谢的影响[J].生态学杂志, 2014, 33(9):2448-2453.
• [4] KOZUKI Y, YAMANAKA R, MATSUSHIGE M, et al. The after-effects of hypoxia exposure on the clam Ruditapes philippinarum in Omaehama beach, Japan[J]. Estuarine,Coastal and Shelf Science, 2013, 116:50-56.
• [5]王彦欣,向蒙,陈磊,等.长期低氧对斑马鱼肝脏糖脂代谢的影响[J].大连海洋大学学报, 2023, 38(3):429-437.
• [6]吴丽娜.低氧预适应与哈维氏弧菌刺激对魁蚶耐低氧的影响[D].上海：上海海洋大学, 2022.
• [7] HE C Z, HAO R J, DENG Y W, et al. Response of pearl oyster Pinctada fucata martensii to allograft-induced stress from lipid metabolism[J]. Fish&Shellfish Immunology,2020, 98:1001-1007.
• [8] YANG C Y, HAO R J, DENG Y W, et al. Effects of protein sources on growth, immunity and antioxidant capacity of juvenile pearl oyster Pinctada fucata martensii[J]. Fish&Shellfish Immunology, 2017, 67:411-418.
• [9] LI J H, YANG C Y, WANG Q H, et al. Growth and survival of host pearl oyster Pinctada fucata martensii(Dunker,1880)treated by different biofouling-clean methods in China[J]. Estuarine, Coastal and Shelf Science, 2018, 207:104-108.
• [10]刘傲东,李力,侯令,等.缺氧与饥饿对马氏珠母贝滤水率和耗氧率的影响[J].基因组学与应用生物学, 2016, 35(3):595-602.
• [11] CHEN J Y, HUANG J, PENG J Q, et al. Effects of hypoxic stress on the digestion, energy metabolism, oxidative stress regulation, and immune function of the pearl oyster(Pinctada fucata martensii)[J]. Aquaculture Reports,2022, 25:101246.
• [12] DIAZ R J, ROSENBERG R. Spreading dead zones and consequences for marine ecosystems[J]. Science, 2008,321(5891):926-929.
• [13] DU X D, FAN G Y, JIAO Y, et al. The pearl oyster Pinctada fucata martensii genome and multi-omic analyses provide insights into biomineralization[J]. GigaScience, 2017, 6(8):1-12.
• [14] HERMES-LIMA M, MOREIRA D C, RIVERA-INGRAHAM G A, et al. Preparation for oxidative stress under hypoxia and metabolic depression:Revisiting the proposal two decades later[J]. Free Radical Biology&Medicine, 2015, 89:1122-1143.
• [15] ZENG Q H, LUO M Z, QIN L R, et al. Effects of hypoxia stress on survival, antioxidant and anaerobic metabolic enzymes, and related gene expression of red swamp crayfish Procambarus clarkii[J]. Biology, 2024, 13(1):33.
• [16] GOSTYUKHINA O L, YU A A, CHELEBIEVA E S, et al.Adaptive potential of the Mediterranean mussel Mytilus galloprovincialis to short-term environmental hypoxia[J].Fish&Shellfish Immunology, 2022, 131:654-661.
• [17] ELMORE S. Apoptosis:a review of programmed cell death[J]. Toxicologic Pathology, 2007, 35(4):495-516.
• [18] GARRIDO C, GALLUZZI L, BRUNET M, et al. Mechanisms of cytochrome c release from mitochondria[J]. Cell Death&Differentiation, 2006, 13(9):1423-1433.
• [19] BARRY P, CHAO Z. The heat shock proteins:their roles as multi-component machines for protein folding[J]. Fungal Biology Reviews, 2009, 22(3):110-119.
• [20] SREEDHAR A S, KALMáR E, CSERMELY P, et al. Hsp90isoforms:functions, expression and clinical importance[J].FEBS Letters, 2004, 562(1/2/3):11-15.
• [21]候利波,陆银月,任秋霖,等.低氧胁迫下中华绒螯蟹血淋巴细胞的转录组学[J].水产学报, 2023, 47(4):27-41.
• [22] ODA K, MATSUOKA Y, FUNAHASHI A, et al. A comprehensive pathway map of epidermal growth factor receptor signaling[J]. Molecular Systems Biology, 2005,1:2005.0010.
• [23] WANG Q H, HAO R J, ZHAO X X, et al. Identification of EGFR in pearl oyster(Pinctada fucata martensii)and correlation analysis of its expression and growth traits[J].Bioscience, Biotechnology, and Biochemistry, 2018, 82(7):1073-1080.
• [24] MUDUMANA S P, HENTSCHEL D, LIU Y, et al. Odd skipped related1 reveals a novel role for endoderm in regulating kidney versus vascular cell fate[J]. Development,2008, 135(20):3355-3367.
• [25] YANG C Y, YANG J M, HAO R J, et al. Molecular characterization of OSR1 in Pinctada fucata martensii and association of allelic variants with growth traits[J].Aquaculture, 2020, 516:734617.
• [26] MASSAGUéJ, CHEIFETZ S, ENDO T, et al. Type beta transforming growth factor is an inhibitor of myogenic differentiation[J]. Proceedings of the National Academy of Sciences of the United States of America, 1986, 83(21):8206-8210.
• [27]杨发明,林海生,秦小明,等.珍珠贝外套膜酶解产物促进皮肤创伤愈合效果研究[J].南方水产科学, 2019, 15(5):92-98.
• [28] KNIGHT P G, GLISTER C. TGF-beta superfamily members and ovarian follicle development[J]. Reproduction, 2006,132(2):191-206.
• [29] MOORE E E, BENDELE A M, THOMPSON D L, et al.Fibroblast growth factor-18 stimulates chondrogenesis and cartilage repair in a rat model of injury-induced osteoarthritis[J]. Osteoarthritis and Cartilage, 2005, 13(7):623-631.
• [30] SHIMOAKA T, OGASAWARA T, YONAMINE A, et al.Regulation of osteoblast, chondrocyte, and osteoclast functions by fibroblast growth factor(FGF)-18 in comparison with FGF-2 and FGF-10[J]. The Journal of Biological Chemistry,2002, 277(9):7493-7500.
• [31] HAO R J, MO C C, ADZIGBLI L, et al. Molecular cloning and polymorphism analysis of Pm FGF18 from Pinctada fucata martensii[J]. Journal of Marine Science and Engineering,2020, 8(11):896.
• [32] HAO R J, ZHENG Z, DU X D, et al. Molecular cloning and characteristics analysis of Pmtgfbr1 from Pinctada fucata martensii[J]. Biotechnology Reports, 2018, 19:e00262.
• [33] YANG C Y, WANG Q H, HAO R J, et al. Effects of replacing microalgae with an artificial diet on pearl production traits and mineralization-related gene expression in pearl oyster Pinctada fucata martensii[J]. Aquaculture Research, 2017, 48(10):5331-5337.
• [34] ZHAO X X, WANG Q H, JIAO Y, et al. Identification of genes potentially related to biomineralization and immunity by transcriptome analysis of pearl sac in pearl oyster Pinctada martensii[J]. Marine Biotechnology, 2012, 14(6):730-739.
• [35] TSUKAMOTO D, SARASHINA I, ENDO K. Structure and expression of an unusually acidic matrix protein of pearl oyster shells[J]. Biochemical and Biophysical Research Communications, 2004, 320(4):1175-1180.
• [36] SUZUKI M, SARUWATARI K, KOGURE T, et al. An acidic matrix protein, Pif, is a key macromolecule for nacre formation[J]. Science, 2009, 325(5946):1388-1390.
• [37] YAN F, JIAO Y, DENG Y W, et al. Tissue inhibitor of metalloproteinase gene from pearl oyster Pinctada martensii participates in nacre formation[J]. Biochemical and Biophysical Research Communications, 2014, 450(1):300-305.
• [38] SUN X J, TU K, LI L, et al. Integrated transcriptome and metabolome analysis reveals molecular responses of the clams to acute hypoxia[J]. Marine Environmental Research,2021, 168:105317.